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Isolation of Functional Cardiac Immune Cells
Published on: December 5, 2011
A novel technique for isolating functional mast cells from the heart
L G Morgan1, S P Levick, T G Voloshenyuk
1Cell and Developmental Biology and Anatomy, School of Medicine, University of South Carolina, Columbia, SC 29208, USA.
Summary
A new method successfully isolated functional cardiac mast cells from rats. This technique allows for better study of mast cells
Area of Science:
- Immunology and Cardiovascular Science
Background:
- Cardiac mast cells play a role in myocardial disease, but their isolation and functional characterization have been challenging.
- Existing isolation techniques for peritoneal and pleural mast cells needed adaptation for cardiac mast cells.
Purpose of the Study:
- To assess the feasibility of adapting mast cell isolation techniques for cardiac mast cells.
- To recover functional cardiac mast cells responsive to secretagogues like compound 48/80.
Main Methods:
- A novel rat protocol involved injecting HBSS into the pericardial space and aspirating to recover epicardial mast cells.
- Cellular functionality was assessed by measuring histamine release in response to compound 48/80, A23187, and substance P.
- Mast cell phenotype was confirmed using immunofluorescence, staining, and Western blotting for chymase and tryptase.
Main Results:
- Isolated cardiac mast cells exhibited low basal histamine release and high responsiveness to secretagogues.
- These mast cells were identified as the connective tissue type, aligning with prior characterizations.
- The novel technique yielded viable and functional mast cells.
Conclusions:
- The pericardial aspiration technique provides a straightforward method for isolating and characterizing cardiac mast cells.
- This advancement enables better in vitro study of epicardial mast cell function.
- Improved understanding of mast cell contributions to myocardial disease is facilitated.
